1. Field of the Invention
The present invention relates to pharmaceutical compositions comprising molecules that are inhibitors of xcex14 mediated (including xcex14xcex27) adhesion and which could be useful in treating conditions such as asthma, diabetes, rheumatoid arthritis, inflammatory bowel disease and other diseases involving leukocyte infiltration of the gastrointestinal tract or other epithelial lined tissues; such as, skin, urinary tract, respiratory airway and joint synovium.
The inhibitors of the present invention could also be useful in treating conditions involving leukocyte infiltration of other tissues including lung, blood vessels, heart and nervous system as well as transplanted organs such as kidney, liver, pancreas and heart.
2. Description of the Related Art
The adhesion of leukocyte to endothelial cells or extracellular matrix proteins is a fundamental process for immunity and inflammation and involves multiple adhesive interactions. The earliest events in this process include leukocyte rolling followed by changes in integrin avidity, which leads to subsequent firm adhesion (for reviews see Butcher, Cell 67:1033-1036 (1991); Harlan, Blood 3:513-525 (1985); Hemler, Annu. Rev. Immunol. 8:365-400 (1990);
Osborn, Cell 62:3-6 (1990); Shimizu et al., Immunol. Rev. 114:109-143 (1990); Springer, Nature 346:425-434 (1990); Springer, Cell 76:301-314 (1994)). In response to chemotactic factors, the leukocytes must migrate through two adjacent endothelial cells and into tissues that are composed, in part, of the extracellular matrix protein fibronectin (FN) (see Wayner et al., J. Cell Biol. 105:1873-1884 (1987)) and collagen (CN) (see Bornstein et al., Ann. Rev. Biochem. 49:957-1003 (1980) and Miller, Chemistry of the collagens and their distribution. In Connective Tissue Biochemistry. K. A. Piez and A. H. Reddi, editors. Elsevier, Amsterdam. 41-78. (1983)) Important recognition molecules that participate in these reactions belong to the integrin gene superfamily (for reviews see Hemler, Annu. Rev. Immunol. 8:365-400 (1990); Hynes, Cell 48:549-554 (1987); Shimizu et al., Immunol. Rev. 114:109-143 (1990); and Springer, Nature 346:425-434 (1990)).
Integrins are composed of non-covalently associated subunits, referred to as the alpha (xcex1) and beta (xcex2) subunits (for reviews see Hemler, Annu. Rev. Immunol. 8:365-400 (1990); Hynes, Cell 48:549-554 (1987); Shimizu et al., Immunol. Rev. 114:109-143 (1990); and Springer, Nature 346:425-434 (1990)). To date, 8 integrin xcex2 subunits have been identified which can associate with 16 distinct xcex1 subunits to form 22 distinct integrins. The xcex27 integrin subunit, first cloned by Erle et al., (Erle et al., J. Biol. Chem. 266:11009-11016 (1991)) is expressed only on leukocytes and is known to associate with two distinct xcex1 subunits, xcex14 (Ruegg et al., J. Cell Biol. 117:179-189 (1992)) and xcex1E (Cerf-Bensussan et al., Eur. J. Immunol. 22:273-277 (1992) and Kilshaw et al., Eur. J. Immunol. as its sole ligand. 
The xcex14xcex27 complex has three known ligands (VCAM, CS-1, MAdCAM). One ligand which shows unique specificity for xcex14xcex27 is Mucosal Addressing Cell Adhesion Molecule (MAdCAM) (see Andrew et al., J. Immunol 153:3847-3861 (1994); Briskin et al., Nature 363:461-464 (1993); and Shyjan et al., J. Immunol 156:2851-2857 (1996)). MAdCAM is highly expressed on Peyer""s patch high endothelial venules, in mesenteric lymph nodes, and on gut lamina propria and mammary gland venules (Berg et al., Immunol. Rev. 105:5 (1989)). Integrin xcex14xcex27 and MAdCAM have been shown to be important in regulating lymphocyte trafficking to normal intestine (Holzmann et al., Cell 56:37 (1989)).
The second ligand for xcex14xcex27 is connecting segment 1 (CS-1), an alternatively spliced region of the FN A chain (see Guan et al., Cell 60:53-61 (1990) and Wayner et al., J. Cell Biol. 109:1321-1330 (1989)). The cell-binding site within this alternatively spliced region is composed of 25 amino acids where the carboxy terminal amino acid residues, EILDVPST, form the recognition motif (see Komoriya et al., J. Biol. Chem. 266:15075-15079 (1991) and Wayner et al., J. Cell Biol. 116:489-497 (1992)).
The third ligand for xcex14xcex27 is vascular cell adhesion molecule 1 (VCAM-1), a cytokine inducible protein expressed on endothelial cells (see Elices et al., Cell 60:577-584 (1990) and Ruegg et al., J. Cell Biol. 117:179-189 (1992)). VCAM and CS-1 (see Elices et al., Cell 60:577-584 (1990)) are two ligands which are shared by xcex14xcex27 and xcex14xcex21. It remains to be unequivocally shown whether MAdCAM, VCAM and CS-1 bind to the same site on xcex14xcex27. Using a panel of monoclonal antibodies, Andrew et al., showed that xcex14xcex27 interaction with its three ligands involve distinct but overlapping epitopes (Andrew et al., J. Immunol 153:3847-3861 (1994)).
A number of in vitro and in vivo studies indicate that xcex14 plays a critical role in the pathogenesis of a variety of diseases. Monoclonal antibodies directed against xcex14 have been tested in a variety of disease models. Efficacy of anti-xcex14 antibody was demonstrated in a rat and mouse model of experimental autoimmune encephalomyelitis (see Baron et al., J. Exp. Med. 177:57-68 (1993) and Yednock et al., Nature 356:63-66(1992)). A significant number of studies have been done to evaluate the role of xcex14 in allergic airways (see Abraham et al., J. Clin. Invest. 93:776-787 (1994); Bochner et al., J. Exp. Med. 173:1553-1556 (1991); Walsh et al., J. Immunol 146:3419-3423 (1991); and Weg et al., J. Exp. Med. 177:561-566 (1993)). For example, monoclonal antibodies to xcex14 were effective in several lung antigen challenge models (see Abraham et al., J. Clin. Invest. 93:776-787 (1994) and Weg et al., J. Exp. Med. 177:561-566 (1993)). Interestingly, blockade of cellular recruitment is not seen in certain lung models even though there is abrogation of the late phase response (see Abraham et al., J. Clin. Invest. 93:776-787 (1994)). The cotton-top tamarin, which experiences spontaneous chronic colitis, showed a significant attenuation of colitis when anti-xcex14 antibody was administered (see Bell et al., J. Immunol. 151:4790-4802 (1993) and Podolsky et al., J. Clin. Invest. 92:372-380 (1993)). Monoclonal antibody to xcex14 inhibits insulitis and delays the onset of diabetes in the non-obese diabetic mouse (see Baron et al., J. Clin. Invest. 93:1700-1708 (1994); Burkly et al., i Diabetes 43:529-534 (1994); and Yang et al., Proc. Natl. Acad. Sci. USA 90:10494-10498 (1993)). Other diseases where xcex14 has been implicated include rheumatoid arthritis (see Laffon et al., J. Clin. Invest. 88:546-552 (1991) and Morales-Ducret et al., J. Immunol. 149:1424-1431 (1992)) and atherosclerosis (see Cybulsky et al., Science 251:788-791 (1991)). Delayed type hypersensitivity reaction (see Issekutz, J. Immunol. 147:4178-4184 (1991)) and contact hypersensitivity response (see Chisholm et al., Eur. J. Immunol. 23:682-688 (1993) and Ferguson et al., J. Immunol. 150:1172-1182 (1993)) are also blocked by anti-xcex14 antibodies. For an excellent review of in vivo studies implicating xcex14 in disease (see Lobb et al., J. Clin. Invest. 94:1722-1728 (1995)).
Although these studies clearly implicate xcex14 in a variety of diseases, it is not clear whether the inhibition seen was due to blocking xcex14xcex21, xcex14xcex27, or both. Recently, several studies have addressed this issue using an antibody which recognizes the xcex14xcex27 complex (see Hesterberg et al., Gastroenterology (1997)), antibodies against xcex27 or antibodies directed against MAdCAM (see Picarella et al., J. Immunol. 158:2099-2106 (1997)), for which xcex14xcex21 does not bind. In the primate model of inflammatory bowel disease, it was shown that antibodies to the xcex14xcex27 complex ameliorated inflammation and decreased diarrhea (see Hesterberg et al., Gastroenterology, 111:1373-1380(1996)). In a second model, monoclonal antibodies to xcex27 or MAdCAM blocked recruitment of lymphocytes to the colon and reduced the severity of inflammation in the colon of scid mice reconstituted with CD45RBhigh CD4+ cells (see Picarella et al., J. Immunol. 158:2099-2106 (1997)). This, together with the fact that gut-associated lymphoid tissue is severely impaired in xcex27 knock out mice, suggests that xcex14xcex27 may be an important intervention point for inflammatory bowel disease.
The expression of xcex14xcex27 on a variety of leukocytes and the increase in xcex14xcex27 positive cells in diseased tissues implicates that the receptor may play an important role in cellular recruitment to other sites of inflammation in addition to trafficking to the gut. CD4+, CD8+ T-cells, B-cells, NK cells, and eosinophils from human peripheral blood were shown to express high levels of xcex14xcex27 (see Picarella et al., J. Immunol. 158:2099-2106 (1997)). Increased numbers of xcex14xcex27+ T-cells were found in the synovial membrane of rheumatoid arthritis patients and it was predicted that the augmented expression of xcex14xcex27 may contribute to the development and perpetuation of this disease (see Lazarovits et al., J. Immunol. 151:6482-6489 (1993)). In the nonobese diabetic mouse, MAdCAM was expressed on high endothelial venules in inflamed islets within the pancreas suggesting a role for xcex14xcex27 in diabetes (see Kelner et al., Science 266:1395-1399 (1994)). The distribution of xcex14xcex27 on lymphocytes and eosinophils (see Erle et al., J. Immunol. 153:517-528 (1994)), together with in vitro studies showing that xcex14xcex27mediates human eosinophil adhesion to VCAM, CS-1 and MAdCAM (see Walsh et al., (Immunology 89:112-119, 1996), suggests that this integrin may be a target molecule in asthma. Collectively, these data suggest that integrin xcex14xcex27 may play an important role in a variety of inflammatory diseases.
N-terminal domain (domain 1) of MAdCAM has homology to the N-terminal integrin recognition domains in both VCAM and ICAM (see Briskin et al., Nature 363:461-464 (1993)). Using site-directed mutagenesis on MAdCAM, the binding motif was identified in the first domain as three linear amino acid residues within a C-D loop (see Viney et al., J. Immunol. 157:2488-2497 (1996)). Mutations of L40, D41 and T42 resulted in a complete loss of binding activity to xcex14xcex27, suggesting that LDT on MADCAM is involved in binding loop (see Viney et al., J. Immunol. 157:2488-2497 (1996)). Alignment of this region on MAdCAM with other integrin ligands such as VCAM or CS-1 reveals that there is a conserved binding motif or consensus sequence, consisting of G/Q I/L E/DT/S and P/S residues (see Briskin et al., J. Immunol. 156:719-726 (1996)). Further support comes from the fact that linear and cyclic peptides containing LDT were shown to block cell adhesion to MAdCAM in vitro (see Shroff et al., Bioorganic and Medicinal Chemistry Letters 6:2495-2500 (1996) and Viney et al., J. Immunol. 157:2488-2497 (1996)).
The use of monoclonal antibodies against integrins in vivo has demonstrated that a number of integrins are indeed valid therapeutic targets for inflammatory and cardiovascular diseases and in organ transplantation. The objective here was to define an orally bioavailable, non-peptide, small molecule antagonist of xcex14xcex27. Small molecules that are potent inhibitors of xcex14xcex27 mediated adhesion to either MAdCAM, VCAM, or CS-1 and which could be useful for the treatment of inflammatory disease are disclosed.
BOP-C1: Bis(2-oxo-3-oxazolidinyl)phosphinic chloride
BOP reagent: Benzotriazol-1-yloxy-tris(dimethylamino)phosphonium hexafluorophosphate
DCC: 1,3-Dicyclohexylcarbodiimide
EDC: 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide
THF: Tetrahydrofuran
DMF: N,N-Dimethylformamide
DIEA: Diisopropylethylamine
DMAP: 4-(N,N-Dimethylamino)pyridine
DBU: 1,8-Diazabicyclo[5.4.0]undec-7-ene
CDI: Carbonyldiimidazole
HOBT: 1-Hydroxybenzotriazole
Boc: tert-Butoxycarbonyl
Tf2O: Triflic anhydride
Tf: Trifluoromethanesulfonyl
TFA: Trifluoroacetic acid
DME: 1,2-Dimethoxyethane
MsCl: Methanesulfonyl chloride
DIAD: Diisopropyl azodicarboxylate
Ac: Acetyl
Me: Methyl
Et: Ethyl
Ph: Phenyl
Bn: Benzyl
EtOAc: Ethyl acetate (=AcOEt)
mCPBA: m-Chloroperbenzoic acid
TMS: Trimethylsilyl
h: hour(s)
min: minute(s)
satd: Saturated
Additionally, several phrases are utilized for which specific meanings and interpretations exist. These are as follows:
The use of xe2x80x9clowerxe2x80x9d preceding a group such as alkyl, alkoxy, alkylene or alkane are meant to encompass 1 to 6 carbon atoms either in a straight chain or in a branched chain and the use of xe2x80x9clowerxe2x80x9d preceding alkanoyl, alkenyl, or alkenylene are meant to encompass 2 to 7 carbon atoms either in a straight chain or in a branched chain. The use of xe2x80x9clowerxe2x80x9d preceding cycloalkyl or cycloalkoxy are meant to encompass 3 to 7 carbon atoms.
The use of phrases such as xe2x80x9cmorpholino-lower alkylxe2x80x9d, xe2x80x9chydroxy-lower alkoxyxe2x80x9d and the like are meant to refer to groups wherein the functional group preceding the hyphen is a substituent of the functional group that follows the hyphen. For example, xe2x80x9chydroxy-lower alkoxyxe2x80x9d would refer to a lower alkoxy group containing at least one hydroxy substituent.
The use of phrases such as xe2x80x9ca lower alkyl group substituted by a halogen atomxe2x80x9d, xe2x80x9cphenyl group substituted by a lower alkoxy groupxe2x80x9d and the like are meant to refer to functional groups containing at least one substituent. For example, xe2x80x9ca lower alkyl group substituted by a halogen atomxe2x80x9d would refer to a lower alkyl group containing at least one halogen atom, and xe2x80x9cphenyl group substituted by a lower alkoxy groupxe2x80x9d would refer to at least one lower alkoxy group. This type of phraseology is meant to be interpreted by one of skill in the art, therefore, any deviations and combinations of this type of nomenclature is also within the abilities of those skilled in the art to interpret. Accordingly, this type of nomenclature is not to be applied to combinations that would not result in a realistic type of molecule or substituent.
The present invention relates to a pharmaceutical composition comprising therapeutically effective amount of a compound of the formula [I]: 
wherein
Ring A is an aromatic hydrocarbon ring or a heterocyclic ring;
Q is a bond, a carbonyl group, a lower alkylene group which may be substituted by a hydroxyl group or phenyl group, a lower alkenylene group, or a xe2x80x94O-(lower alkylene)-group;
n is an integer of 0, 1 or 2;
W is oxygen atom, sulfur atom, a xe2x80x94CHxe2x95x90CHxe2x80x94 group or a xe2x80x94Nxe2x95x90CHxe2x80x94 group;
Z is oxygen atom or sulfur atom;
R1, R2, R3 are the same or different and are selected from the group consisting of:
a) hydrogen atom,
b) a halogen atom,
c) a substituted or unsubstituted lower alkyl group,
d) a substituted or unsubstituted lower alkoxy group,
e) a nitro group,
f) a substituted or unsubstituted amino group,
g) a carboxyl group or an amide or an ester thereof,
h) a cyano group,
i) a lower alkylthio group,
j) a lower alkanesulfonyl group,
k) a substituted or unsubstituted sulfamoyl group,
l) a substituted or unsubstituted aryl group,
m) a substituted or unsubstituted heterocyclic group, and
n) hydroxyl group;
or two of R1, R2 and R3 may combine each other at the terminal thereof to form a lower alkylenedioxy group;
R4 is tetrazolyl group, a carboxyl group, or an amide or an ester thereof;
R5 is a group selected from the group consisting of:
a) a hydrogen atom,
b) a nitro group,
c) a substituted or unsubstituted amino group,
d) a hydroxyl group,
e) a lower alkanoyl group,
f) a substituted or unsubstituted lower alkyl group,
g) a lower alkoxy group,
h) a halogen atom, and
i) 2-oxopyrrolidinyl group;
R6 is a group selected from the group consisting of:
a) a substituted or unsubstituted phenyl group, and
b) a substituted or unsubstituted heteroaryl group; or a pharmaceutically acceptable salt thereof.
The present invention also relates to a method for treating or preventing conditions caused by xcex14 (including xcex14xcex27 and xcex14xcex21) mediated cell adhesion which comprises administering a compound of the formula [I].
Further, the present invention also relates to a novel compound, which is a compound of the formula [I] with the proviso that when Ring A is a benzene ring, it is not substituted with methyl group in the 3- and the 5-positions or in the 2- and the 4-positions; or a pharmaceutically acceptable salt thereof.
The novel compound of the present invention may exist in the form of optical isomers based on asymmetric carbon atoms thereof, and the present invention also includes these optical isomers and mixtures thereof.
In an embodiment of the present invention, the steric configuration of the compound need not be fixed. The compound of the present invention may be a compound with a sole configuration or a mixture thereof with several different configurations.
In the above formula (I), xe2x80x9caromatic hydrocarbon ringxe2x80x9d may be a mono-, bi- or tri-cyclic aromatic hydrocarbon ring such as a benzene ring, a naphthalene ring, an anthracene ring, a fluorene ring.
In the above formula (I), xe2x80x9cheterocyclic ringxe2x80x9d may be a heteroatom-containing mono-, bi- or tri-cyclic ring. Examples of xe2x80x9cheterocyclic ringxe2x80x9d may be pyridine ring, pyrimidine ring, pyridazine ring, pyrazine ring, quinoline ring, isoquinoline ring, quinazoline ring, phthalazine ring, imidazole ring, isoxazole ring, pyrazole ring, oxazole ring, thiazole ring, indole ring, benzazole ring, benzothiazole ring, benzimidazole ring, benzofuran ring, furan ring, thiophene ring, pyrrole ring, oxadiazole ring, thiadiazole ring, triazole ring, tetrazole ring, pyrrole ring, indoline ring, indazole ring, isoindole ring, purine ring, morpholine ring, quinoxaline ring, benzothiophene ring, pyrrolidine ring, benzofurazane ring, benzothiadiazole ring, thiazolidine ring, imidazothiazole ring, dibenzofuran ring, and isothiazole ring.
In the above formula (I), xe2x80x9caryl groupxe2x80x9d may be a mono-, bi- or tri-cyclic aromatic group. Examples of xe2x80x9caryl groupxe2x80x9d may be a phenyl group, a naphthyl group, an anthryl group and a fluorenyl ring.
In the above formula (I), xe2x80x9cheterocyclic groupxe2x80x9d may be a mono-, bi- or tri-cyclic ring containing a heteroatom such as nitrogen atom, oxygen atom, and sulfur atom. Examples of xe2x80x9cheterocyclic groupxe2x80x9d may be pyridyl group, pyrimidinyl group, pyridazinyl group, pyrazinyl group, quinolyl group, isoquinolyl group, quinazolinyl group, phthalazinyl group, imidazolyl group, isoxazolyl group, pyrazolyl group, oxazolyl group, thiazolyl group, indolyl group, benzazolyl group, benzothiazolyl group, benzimidazolyl group, benzofuranyl group, furyl group, thienyl group, pyrrolyl group, oxadiazolyl group, thiadiazolyl group, triazolyl group, tetrazolyl group, pyrrolyl group, indolinyl group, indazolyl group, isoindolyl group, purinyl group, morpholinyl group, quinoxalinyl group, benzothienyl group, pyrrolidinyl group, benzofurazanyl group, benzothiadiazolyl group, thiazolidinyl group, imidazothiazolyl group, dibenzofuranyl group, isothiazolyl group, pyrrolinyl group, piperidinyl group, piperazinyl group, and tetrahydropyranyl group.
In the above formula (I), xe2x80x9cheteroaryl groupxe2x80x9d may be a mono-, bi- or tri-cyclic aromatic group containing a heteroatom such as nitrogen atom, oxygen atom, and sulfur atom. Examples of xe2x80x9cheteroaryl groupxe2x80x9d may be a xe2x80x9cheterocyclic ringxe2x80x9d other than pyrrolidinyl group, pyrrolinyl group, piperidinyl group, piperazinyl group, morpholinyl group, and tetrahydropyranyl group. Preferable examples of the xe2x80x9cheteroaryl groupxe2x80x9d may be pyridyl group, thienyl group, benzofuranyl group, pyrimidyl group, and isoxazolyl group.
The novel compound among the compound [I] of the present invention is indicated as follows: 
wherein
Ring A is an aromatic hydrocarbon ring or a heterocyclic ring;
Q is a bond, a carbonyl group, a lower alkylene group which may be substituted by a hydroxyl group or phenyl group, a lower alkenylene group, or a xe2x80x94O-(lower alkylene)-group;
n is an integer of 0, 1 or 2;
W is oxygen atom, sulfur atom, a xe2x80x94CHxe2x95x90CHxe2x80x94 group or a xe2x80x94Nxe2x95x90CHxe2x80x94 group;
Z is oxygen atom or sulfur atom;
R1, R2 and R3 are the same or different and are selected from the group consisting of:
a) hydrogen atom,
b) a halogen atom,
c) a substituted or unsubstituted lower alkyl group,
d) a substituted or unsubstituted lower alkoxy group,
e) a nitro group,
f) a substituted or unsubstituted amino group,
g) a carboxyl group or an amide or an ester thereof,
h) a cyano group,
i) a lower alkylthio group,
j) a lower alkanesulfonyl group,
k) a substituted or unsubstituted sulfamoyl group,
l) a substituted or unsubstituted aryl group,
m) a substituted or unsubstituted heterocyclic group, and
n) hydroxyl group;
or two of R1, R2 and R3 may combine each other at the terminal thereof to form a lower alkylenedioxy group;
R4 is tetrazolyl group, a carboxyl group, or an amide or an ester thereof;
R5 is a group selected from the group consisting of:
a) a hydrogen atom,
b) a nitro group,
c) a substituted or unsubstituted amino group,
d) a hydroxyl group,
e) a lower alkanoyl group,
f) a substituted or unsubstituted lower alkyl group,
g) a lower alkoxy group,
h) a halogen atom, and
j) 2-oxopyrrolidinyl group;
R6 is a group selected from the group consisting of:
a) a substituted or unsubstituted phenyl group,
b) a substituted or unsubstituted heteroaryl group;
with the proviso that when Ring A is a benzene ring, the ring is not substituted with methyl group in the 3- and the 5-positions or in the 2- and the 4-positions; or a pharmaceutically acceptable salt thereof.
A preferred configuration of the active ingredient of the present invention is represented by the formula [I-A]: 
wherein symbols are the same as defined above.
A preferred embodiment of the present invention is the compound [I] with the additional proviso that when Ring A is a benzene ring, the ring is substituted in at least one of 2- and 6-positions.
Another preferred embodiment of the present invention is the compound (I) wherein R1, R2 and R3 are selected from the group consisting of:
a) hydrogen atom,
b) a halogen atom,
c) a substituted or unsubstituted lower alkoxy group,
d) a nitro group,
e) a substituted or unsubstituted amino group,
f) a carboxyl group or an amide or an ester thereof,
g) a cyano group,
h) a lower alkylthio group,
i) a lower alkanesulfonyl group,
j) a substituted or unsubstituted sulfamoyl group,
k) a substituted or unsubstituted aryl group,
l) a substituted or unsubstituted heterocyclic group, and
m) hydroxyl group, or two of R1, R2 and R3 may combine with each other at the terminal thereof to form a lower alkylenedioxy group.
A more preferred configuration of the active ingredient of the present invention is represented by the formula [I-B]: 
wherein symbols are the same as defined above.
In more preferred embodiment of the present invention,
R1 is hydrogen atom, a halogen atom, carboxyl group, carbamoyl group, nitro group, a substituted or unsubstituted amino group, a substituted or unsubstituted heterocyclic ring;
R2 is hydrogen atom, a lower alkyl group or a halogen atom;
R3 is hydrogen atom, a lower alkyl group or a halogen atom;
R6 is a phenyl group which may be substituted at 2-, 4-, and/or 6-position of the phenyl group by a group selected from the group consisting of:
1) a halogen atom,
2) a substituted or unsubstituted lower alkoxy group,
3) a substituted or unsubstituted lower alkyl group,
4) a substituted or unsubstituted amino group,
5) a substituted or unsubstituted carbamoyl group, and
6) a substituted or unsubstituted sulfamoyl group.
In further preferred embodiment of the present invention, R6 is a phenyl group which may be substituted by a group selected from the group consisting of:
1) a lower alkoxy group, and
2) a lower alkyl group which may be substituted by a group selected from a substituted or unsubstituted amino group, a substituted or unsubstituted piperidinyl group, a substituted or unsubstituted morpholino group, a substituted or unsubstituted piperazinyl group, a substituted or unsubstituted pyrrolidinyl group, and a substituted or unsubstituted imidazolidinyl group.
In another embodiment of the present invention,
Ring A is a benzene ring, a pyridine ring, a pyrazine ring, a furan ring, an isoxazole ring, a benzofuran ring, a thiophene ring, a pyrrole ring, or an indole ring;
R1, R2 and R3 are selected from the group consisting of:
a) hydrogen atom,
b) a halogen atom,
c) a lower alkyl group which may be substituted by a halogen atom or a (halogenobenzoyl)amino group,
d) a lower alkoxy group which may be substituted by a halogen atom,
e) a nitro group,
f) an amino group which may be substituted by 1-2 groups selected from the group consisting of 1) a lower alkyl group, 2) a lower alkanoyl group, 3) a halogenobenzoyl group, 4) a lower alkoxycarbonyl group, 5) a lower alkanesulfonyl group which may be substituted by a halogen atom, 6) a benzenesulfonyl group which may be substituted by a lower alkyl group, a trihalogeno-lower alkyl group, a halogen atom or a lower alkoxy group, 7) thiophenesulfonyl group, 8) a carbamoyl group which may be substituted by a lower alkyl group, a lower alkyl-phenyl group, 9) a thiocarbamoyl group which may be substituted by a lower alkyl group, phenyl group, a phenyl-lower alkyl group, 10) thiazolinyl group, and 11) a sulfamoyl group which may be substituted by a lower alkyl group;
g) a carboxyl group,
h) a carbamoyl group which may be substituted by a lower alkanesulfonyl group,
i) a lower alkoxycarbonyl group,
j) a cyano group,
k) a lower alkylthio group,
l) a lower alkanesulfonyl group,
m) a sulfamoyl group,
n) a phenyl group,
o) a pyrrolidinyl group which may be substituted by oxo group,
p) a pyrrolyl group which may be substituted by a group selected from the group consisting of 1) a lower alkanoyl group which may be substituted by a halogen atom, 2) a halogen atom, 3) formyl group, and 4) a lower alkyl group which may be substituted by hydroxy group,
q) a thienyl group,
r) an isoxazolyl group which may be substituted by a lower alkyl group,
s) a thiazolyl group,
t) a pyrazolyl group,
u) a pyrazinyl group,
v) a pyridyl group, and
w) hydroxyl group;
R4 is selected from the group consisting of:
a) carboxyl group,
b) a lower alkoxycarbonyl group which may be substituted by 1) pyridyl group or 2) an amino group which may be substituted by a lower alkyl group,
c) a lower cycloalkoxy carbonyl group,
d) a carbamoyl group which may be substituted by a hydroxy group or a lower alkanesulfonyl group, and
e) a tetrazolyl group;
R5 is selected from the group consisting of:
a) a hydrogen atom,
b) a nitro group,
c) an amino group which may be substituted by a lower alkanoyl group, a lower alkoxycarbonyl group or a lower alkanesulfonyl group,
d) a hydroxyl group,
e) a lower alkanoyl group,
f) a lower alkyl group which may be substituted by 1) hydroxyl group, or 2) an imino group which is substituted by hydroxyl group or a lower alkoxy group,
g) a lower alkoxy group,
h) a halogen atom,
i) 2-oxopyrrolidinyl group;
R6 is the group selected from the group consisting of:
a) a phenyl group which may have 1-5 substituents selected from the group consisting of:
1) a halogen atom,
2) a nitro group,
3) a formyl group,
4) a hydroxyl group,
5) a carboxyl group,
6) a lower alkoxy group which may be substituted by a group selected from the group consisting of i) a carboxyl group or an amide or an ester thereof, ii) hydroxyl group, iii) a cyano group, iv) a halogen atom, v) an amino group which may be substituted by a lower alkyl group, vi) a pyridyl group, vii) a thiazolyl group which may be substituted by a lower alkyl group, viii) an isoxazolyl group which may be substituted by a lower alkyl group, ix) a piperidyl group which may be substituted by a lower alkyl group, x) a pyrrolidinyl group which may be substituted by a lower alkyl group, xi) a phenyl group which may be substituted by a halogen atom, xii) a furyl group, xiii) a thienyl group, and xiv) a lower alkoxy group
7) a lower alkyl group which may be substituted by a group selected from the group consisting of i) a halogen atom, ii) hydroxyl group, iii) carboxyl group or an amide or an ester thereof, iv) a lower alkoxy group, v) an amino group which may be substituted by 1-2 groups selected from the group consisting of a lower alkyl group, a hydroxy-lower alkyl group, a (lower alkylamino)-lower alkyl group, phenyl-lower alkyl group, a phenyl group, and a pyridyl group, vi) a piperidinyl group which may be substituted by a lower alkylenedioxy group, an oxo group or a hydroxy group, vii) a morpholino group which may be substituted by a lower alkyl group, viii) thiomorpholino group which may be oxidized, ix) piperazinyl group which may be substituted by a lower alkyl group, a hydroxy-lower alkyl group, a lower alkanoyl group or a phenyl-lower alkyl group, x) pyrrolidinyl group which may be substituted by oxo group, and xi) a imidazolidine group which may be substituted by 1-3 groups selected from the group consisting of a lower alkyl group and oxo group,
8) a lower alkenyl group which may be substituted by carboxyl group or an amide or an ester thereof,
9) an amino group which may be substituted by a group selected from the group consisting of i) a phenyl group, ii) a lower alkoxycarbonyl group, iii) a lower alkanesulfonyl group, iv) a carbamoyl group which may be substituted by a lower alkyl group or a lower alkyl-phenyl group, v) a lower alkanoyl group, vi) a lower alkyl group, vii) a lower alkenyl group, and viii) a thiocarbamoyl group which may be substituted by a lower alkyl group,
10) a carbamoyl group which may be substituted by a lower alkyl group, a hydroxy-lower alkyl group, a morpholino-lower alkyl group, a phenyl-lower alkyl group or a lower alkanesulfonyl group,
11) a sulfamoyl group which may be substituted by a group consisting of i) a lower alkyl group, ii) a benzoyl group, iii) a lower alkoxycarbonyl group, and iv) a lower alkanoyl group,
12) a lower alkenyloxy group,
13) a lower alkylenedioxy group,
14) a piperazinylcarbonyl group which may be substituted by a lower alkyl group,
15) a lower alkanoyl group,
16) cyano group,
17) a lower alkylthio group,
18) a lower alkanesulfonyl group,
19) a lower alkylsulfinyl group, and
20) a group of the formula: xe2x80x94(CH2)qxe2x80x94Oxe2x80x94
wherein q is an integer of 2 or 3;
b) a pyridyl group which may be substituted by a lower alkyl group;
c) a thienyl group which may be substituted by a group selected from the group consisting of:
1) a halogen atom,
2) a lower alkyl group which may be substituted by hydroxyl group,
3) cyano group,
4) formyl group,
5) a lower alkoxy group, and
6) a lower alkanoyl group;
d) a benzofuranyl group;
e) a pyrimidinyl group which may be substituted by a lower alkoxy group;
f) a isoxazolyl group which may be substituted by a lower alkyl group; and
g) a pyrrolyl group which may be substituted by a lower alkoxycarbonyl group.
In preferred embodiment of the present invention,
Ring A is a benzene ring,
Q is a bond,
W is a xe2x80x94CHxe2x95x90CHxe2x80x94 group,
R1 is selected from the group consisting of:
a) hydrogen atom,
b) a halogen atom,
c) a lower alkyl group,
d) a lower alkoxy group,
e) nitro group,
f) an amino group which may be substituted by a group selected from the group consisting of 1) a lower alkyl group, 2) a lower alkanoyl group, 3) a lower alkoxycarbonyl group, 4) a lower alkanesulfonyl group which may be substituted by a halogen atom, 5) a benzenesulfonyl group which may be substituted by a lower alkyl group, a trihalogeno-lower alkyl group, a halogen atom or a lower alkoxy group, 6) thiophenesulfonyl group, 7) a carbamoyl group which may be substituted by a lower alkyl group or a lower alkyl-phenyl group, 8) a thiocarbamoyl group which may be substituted by a lower alkyl group, and 9) a sulfamoyl group which may be substituted by a lower alkyl group,
g) carboxyl group
h) a carbamoyl group which may be substituted by a lower alkanesulfonyl group,
i) a lower alkanesulfonyl group,
j) a sulfamoyl group,
k) phenyl group,
l) a pyrrolidinyl group which may be substituted by oxo group,
l) a pyrrolyl group which may be substituted by a lower alkyl group,
m) a thienyl group,
n) an isoxazolyl group which may be substituted by a lower alkyl group,
o) a thiazolyl group
p) a pyrazolyl group,
q) a pyrazinyl group,
r) a pyridyl group, and
s) hydroxyl group;
R2 is hydrogen atom, or a halogen atom;
R3 is hydrogen atom, or a halogen atom;
R4 is a) a carboxyl group,
b) a lower alkoxycarbonyl group which may be substituted by a lower alkyl-amino group, or
c) a carbamoyl group which may be substituted by a lower alkanesulfonyl group;
R5 is selected from the group consisting of:
a) hydrogen atom,
b) an amino group which may be substituted by a lower alkanoyl group, a lower alkoxycarbonyl group or a lower alkanesulfonyl group,
c) a lower alkanoyl group,
d) a lower alkyl group which may be substituted by 1) hydroxyl group, or 2) an imino group which is substituted by hydroxyl group or a lower alkoxy group,
e) a lower alkoxy group, and
f) a halogen atom;
R6 is a phenyl group which may have 1-5 substituents selected from the group consisting of:
a) a halogen atom,
b) a formyl group,
c) a hydroxyl group,
d) a lower alkoxy group which may be substituted by 1) a carboxyl group, 2) a hydroxyl group, 3) a cyano group, 4) a halogen atom, 5) an amino group which may be substituted by a lower alkyl group, 6) a pyridyl group, 7) a phenyl group, 8) a thienyl group, or 9) a lower alkoxy group,
e) a lower alkyl group which may be substituted by 1) an amino group which may be substituted by a lower alkyl group, a hydroxy-lower alkyl group, a lower alkylamino-lower alkyl group or a phenyl group, 2) a piperidinyl group which may be substituted by a lower alkylenedioxy group, 3) a morpholino group which may be substituted by a lower alkyl group, 4) a thiomorpholino group in which sulfur atom may be oxidized, 5) a piperazinyl group which may be substituted by a lower alkyl group, a hydroxy-lower alkyl group, a lower alkanoyl group or a phenyl-lower alkyl group, 6) pyrrolidinyl group which may be substituted by oxo group, or 7) an imidazolidinyl group which may be substituted by 1-3 groups selected from the group consisting of a lower alkyl group and oxo group,
f) an amino group which may be substituted by 1) a lower alkoxycarbonyl group, 2) a lower alkanesulfonyl group, 3) a carbamoyl group which may be substituted by a lower alkyl group a lower alkyl-phenyl group, 4) a lower alkanoyl group, 5) a lower alkyl group, 6) a lower alkenyl group, or 7) a thiocarbamoyl group which may be substituted by a lower alkyl group,
g) a carbamoyl group which may be substituted by 1) a lower alkyl group, 2) a hydroxy-lower alkyl group, 3) a morpholino-lower alkyl group, 4) a phenyl-lower alkyl group, or 5) a lower alkanesulfonyl group,
h) a sulfamoyl group which may be substituted by a lower alkyl group,
i) a lower alkenyloxy group,
j) a lower alkylenedioxy group,
k) a cyano group,
l) a lower alkylthio group, and
m) a lower alkanesulfonyl group.
In more preferred embodiment of the present invention, R1 is 1) hydrogen atom, 2) a halogen atom, 3) a lower alkanoylamino group, 4) a lower alkoxycarbonylamino group, 5) a lower alkanesulfonylamino group which may be substituted by a halogen atom, 6) a benzenesulfonylamino group which may be substituted by a lower alkyl group, a trihalogeno-lower alkyl group, a halogen atom or a lower alkoxy group, 7) thiophenesulfonylamino group, 8) an ureido group which may be substituted by a lower alkyl group or a lower alkyl-phenyl group, 9) a lower alkyl-thioureido group, or 10) a lower alkylsulfamoylamino group, R2 is a halogen atom, R3 is hydrogen atom or a halogen atom, and R6 is a phenyl group which may have 1-3 substituents selected from the group consisting of 1) a lower alkoxy group, 2) a lower alkyl group which may be substituted by a group selected from the group consisting of a lower alkylamino group, a hydroxy-lower alkylamino group, a lower alkylamino-lower alkylamino group, piperidinyl group, a lower alkyl-piperidinyl group, morpholino group, a lower alkyl-morpholino group, a thiomorpholino group, piperazinyl group, a lower alkyl-piperazinyl group, a lower alkanoyl-piperazinyl group, and a pyrrolidinyl group, 3) a sulfamoyl group which may be substituted by a lower alkyl group, 4) a carbamoyl group which may be substituted by a lower alkyl group.
In another more preferred embodiment of the present invention, R1 is hydrogen atom, R3 is a halogen atom, and R6 is 2-(lower alkoxy)phenyl group, 2,6-di(lower alkoxy)phenyl group, 2,6-di(lower alkoxy)-4-[[N,N-di(lower alkyl)amino]lower alkyl]phenyl group, 2,6-di(lower alkoxy)4-[(4-lower alkyl-1-piperazinyl)lower alkyl]phenyl group, 2,6-di(lower alkoxy)-4-[1-piperidinyl-lower alkyl]phenyl group, 2,6-di(lower alkoxy)-4-[N,N-di(lower alkyl)carbamoyl]phenyl group or 2,6-di(lower alkoxy)-4-[(morpholino)lower alkyl]phenyl group.
In another more preferred embodiment of the present invention, a lower alkoxy group is methoxy group.
Preferred compounds as the active ingredient of the present invention may be selected from the group consisting of:
N-(2,6-dichlorobenzoyl)-4-(2,6-dimethoxyphenyl)-L-phenylalanine;
N-(2,6-dichlorobenzoyl)-4-[2,6-dimethoxy-4-(piperidinomethyl)phenyl]-L-phenylalanine;
N-(2,6-dichlorobenzoyl)-4-[2,6-dimethoxy-4-[(4-methylpiperazinyl)amino]phenyl]-L-phenylalanine;
N-(2,6-dichlorobenzoyl)-4-[2,6-dimethoxy-4-(morpholinomethyl)phenyl]-L-phenylalanine;
N-(2,6-dichlorobenzoyl)-4-[2,6-dimethoxy-4-(N,N-dimethylamino)phenyl]-L-phenylalanine;
N-(2,6-dichlorobenzoyl)-4-[2,6-dimethoxy-4-(N,N-dimethylcarbamoyl)phenyl]-L-phenylalanine;
N-(2,6-dichloro-4-hydroxybenzoyl)-4-(2,6-dimethoxyphenyl)-L-phenylalanine;
N-(2,6-dichlorobenzoyl)-4-(2-ethoxy-6-methoxyphenyl)-L-phenylalanine;
N-(2,6-difluorobenzoyl)-4-(2-6,dimethoxyphenyl)-L-phenylalanine;
N-(2,6-dichlorobenzoyl)-4-(2,3-methylenedioxy-6-methoxyphenyl)-L-phenylalanine;
N-(2,6-dichlorobenzoyl)-3-(1-hydroxyethy)-4-(2,6-dimethoxyphenyl)-L-phenylalanine;
N-(2,6-dichlorobenzoyl)-4-(2,4,6-trimethoxyphenyl)-L-phenylalanine;
N-[2,6-dichloro-4-[(trifluoromethanesulfonyl)amino]benzoyl]-4-(2,6-dimethoxyphenyl)-L-phenylalanine; or
N-[2,6-dichloro-4-[(2-thienylsulfonyl)amino]benzoyl]-4-(2,6-dimethoxyphenyl)-L-phenylalanine;
or a lower alkyl ester such as ethyl ester thereof;
or pharmaceutically acceptable salt thereof.
The active ingredient of the present invention may be used in the form of an ester or amide thereof. As the ester thereof, there may be mentioned a) a lower alkyl ester which may be substituted by 1) pyridyl group, 2) an amino group which may be substituted by a lower alkyl group, 3) a lower alkanoyloxy group, 4) an aryl group; b) a lower alkenyl ester; c) a lower alkynyl ester; d) a lower cycloalkyl ester; e) an aryl ester. As the amide thereof, there may be mentioned an amide (xe2x80x94CONH2) which may be substituted by 1) a lower alkyl group, a lower cycloalkyl group, aryl group, aryl-lower alkyl group, hydroxy group or a lower alkanesulfonyl group;
An ester of the formula [I] includes, for example, an ester which can be converted to the corresponding carboxylic acid in a body, for example, a lower alkyl ester (e.g., methyl ester), a lower alkanoyloxy-lower alkyl ester (e.g., acetoxymethyl ester) and the like. An amide of the formula [I] includes, for example, an N-unsubstituted amide, an N-monosubstituted amide (e.g., an N-lower alkyl amide), an N,N-disubstituted amide (e.g., an N,N-(lower alkyl)(lower alkyl) amide) and the like.
A pharmaceutically acceptable salt of the formula [I] includes, for example, a salt with an inorganic acid (e.g., hydrochloride, sulfate), a salt with an organic acid (e.g., p-toluenesulfonate, maleate), a salt with an inorganic base (e.g., a salt with an alkali metal such as a sodium salt or a potassium salt) or a salt with an amine (e.g., an ammonium salt).
The active ingredient of the present invention may be used either in a free form or in the form of pharmaceutically acceptable salts thereof. Pharmaceutically acceptable salts include acid-addition salts with inorganic acid or organic acid (e.g., hydrochloride, sulfate, nitrate, hydrobromide, methanesulfonate, p-toluenesulfonate, acetate), salt with inorganic base, organic base or amino acid (e.g., triethylamine salt, a salt with lysine, an alkali metal salt, an alkali earth metal, salt and the like).
The active ingredient may be formulated into a pharmaceutical composition comprising a therapeutically effective amount of the compound as defined above and a pharmaceutically acceptable carrier or diluent.
The composition can be used for treating or preventing xcex14 (including xcex14xcex21 and xcex14xcex27 adhesion mediated conditions in a mammal such as a human, especially used for treatment or prevention of xcex14xcex27 adhesion mediated conditions. This method may comprise administering to a mammal or a human patient an effective amount of the compound or composition as explained above.
This method can be used to treat or prevent such inflammatory conditions as rheumatoid arthritis, asthma, psoriasis, eczema, contact dermatitis and other skin inflammatory diseases, diabetes, multiple sclerosis, systemic lupus erythematosus (SLE), inflammatory bowel disease including ulcerative colitis and Crohn""s disease, and other diseases involving leukocyte infiltration of the gastrointestinal tract, or other epithelial lined tissues, such as skin, urinary tract, respiratory airway, and joint synovium. The method can be preferably used for treatment or prevention of inflammatory bowel disease including ulcerative colitis and Crohn""s disease.
The present invention also relates to a method for inhibiting the interaction of a cell bearing a ligand of MAdCAM-1, including xcex14xcex27 integrins, with MAdCAM-1 or a portion thereof (e.g., the extracellular domain), comprising contacting the cell with an active ingredient of the present invention. In one embodiment, the present invention relates to a method of inhibiting the MAdCAM-mediated interaction of a first cell bearing an xcex14xcex27 integrin with MAdCAM, for example with a second cell bearing MAdCAM, comprising contacting the first cell with an active ingredient of the present invention. In another embodiment, the invention relates to a method of treating an individual suffering from a disease associated with leukocyte recruitment to tissues (e.g., endothelium) expressing the molecular MAdCAM-1.
Another embodiment of the present invention is a method of treating an individual suffering from a disease associated with leukocyte infiltration of tissues expressing the molecule MAdCAM-1.
According to the present method, the cell bearing the ligand for MAdCAM-1 is contacted with an effective amount of an (i.e., one or more) inhibitor as represented by Structural Formula [I]. As used herein, an inhibitor is a compound which inhibits (reduces or prevents) the binding of MAdCAM-1 to a ligand, including xcex14xcex27 integrin, and/or which inhibits the triggering of a cellular response mediated by the ligand. An effective amount can be an inhibitory amount (such an amount sufficient to achieve inhibition of adhesion of a cell bearing a MAdCAM-1 ligand to MAdCAM-1). Ligands for MAdCAM-1 include xcex14xcex27 integrins, such as human xcex14xcex27 integrin, and its homologs from other species such as mice (also referred to as xcex14xcex2p or LPAM-1 in mice).
For example, the adhesion of a cell which naturally expresses a ligand for MAdCAM-1, such as a leukocyte (e.g., B lymphocyte, T lymphocyte) or other cells which express a ligand for MAdCAM-1 (e.g., a recombinant cell), to MAdCAM-1 can be inhibited in vitro and/or in vivo according to the present method.
In another aspect, the present invention relates to a method of treating an individual (e.g., a mammal, such as a human or other primate) suffering from a disease associated with leukocyte (e.g., lymphocyte, monocyte) infiltration of tissues (including recruitment and/or accumulation of leukocytes in tissues) which express the molecule MAdCAM-1. The method comprises administering to the individual a therapeutically effective amount of an inhibitor (i.e., one or more inhibitors) of Structural Formula [I]. For example, inflammatory diseases, including diseases which are associated with leukocyte infiltration of the gastrointestinal tract (including gut-associated endothelium), other mucosal tissues, or tissues expressing the molecular MAdCAM-1 (e.g., gut-associated tissues, such as venules of the lamina propria of the small and large intestine; and mammary gland (e.g., lactating mammary gland)), can be treated according to the present method. Similarly, an individual suffering from a disease associated with leukocyte infiltration of tissues as a result of binding of leukocytes to cells (e.g., endothelial cells) expressing the molecule MAdCAM-1 can be treated according to the present invention.
Diseases which can be treated accordingly include inflammatory bowel disease (IBD), such as ulcerative colitis, Crohn""s disease and pouchitis resulting after proctocolectomy and ileoanal anastomosis after IBD; and other gastrointestinal diseases associated with leukocyte infiltration, such as Celiac disease, nontropical Sprue, enteropathy associated with seronegative arthropathies, lymphocytic and graft versus host diseases.
Pancreatitis and insulin-dependent diabetes mellitus are other diseases which can be treated using the present method. It has been reported that MAdCAM-1 is expressed by some vessels in the exocrine pancreas from NOD (nonobese diabetic) mice, as well as from BALB/c and SJL mice. Expression of MAdCAM-1 was reportedly induced on endothelium in inflamed islets of the pancreas of the NOD mouse, and MAdCAM-1 was the predominant address in expressed by NOD islet endothelium at early stages of insulitis (Hanninen, A. et al., J. Clin. Invest., 92: 2509-2515 (1993)). Further, accumulation of lymphocytes expressing xcex14xcex27 within islets was observed, and MAdCAM-1 was implicated in the binding of lymphoma cells via xcex14xcex27 to vessels from inflamed islets (Hanninen, A., et al., J. Clin. Invest., 92: 2509-2515 (1993)).
Examples of inflammatory diseases associated with mucosal tissues which can be treated according to the present method include mastitis (mammary gland), cholecystitis, cholangitis or pericholangitis (bile duct and surrounding tissue of the liver), chronic bronchitis, chronic sinusitis, asthma, and graft versus host disease (e.g., in the gastrointestinal tract). Chronic inflammatory diseases of the lung which result in interstitial fibrosis, such as hypersensitivity pneumonitis, collagen disease (in SLE and RA), sarcoidosis, and other idiopathic conditions can be amenable to treatment.
Vascular cell adhesion molecule-1 (VCAM-1), which recognizes the xcex14xcex21 integrin (VLA-4), has been reported to play a role in in vivo leukocyte recruitment (Silber et al., J. Clin. Invest. 93:1554-1563 (1994)). However, these therapeutic targets are likely to be involved in inflammatory processes in multiple organs, and a functional blockade could cause systemic immune dysfunction. In contrast to VCAM-1, MAdCAM-1 is preferentially expressed in the gastrointestinal tract and mucosal tissues, binds the xcex14xcex27 integrin found on lymphocytes, and participates in the homing of these cells to mucosal sites, such as Peyer""s patches in the intestinal wall (Hamann et al., J. Immunol., 152:3282-3293 (1994)). As inhibitors of the binding of MAdCAM-1 to xcex14xcex27 integrin, the active ingredients of the present invention have the potential for fewer side effects due to, for example, effects on other tissue types where adhesion is mediated by other receptors, such as xcex14xcex21 integrin.
Undesired symptoms of the condition listed herein can alleviated using the present method. The symptoms may be caused by inappropriate cell adhesion and/or cell activation to release proinflammatory mediators mediated by xcex14xcex27 integrins. Such inappropriate cell adhesion or signal transduction would typically be expected to occur as a result of increased VCAM and/or MAdCAM expression on the surface of endothelial cells. Increased VCAM, MAdCAM and/or CS-1 expression can be due to a normal inflammatory response or due to abnormal inflammatory states
The present method can be used to assess the inhibitory effect of a compound of the present invention and of other potential antagonists useful in the method on the interaction of MAdCAM-1 with a ligand for MAdCAM-1 in vitro or in vivo.
Compounds suitable for use in therapy can also be evaluated in vivo, using suitable animal models. Suitable animal models of inflammation have been described. For example, NOD mice provide animal model of insulin-dependent diabetes mellitus. CD45 RBHi SCID model provide a model in mice with similarity to both Crohn""s disease and ulcerative colitis (Powrie, F. et al., Immunity, 1: 553-562 (1994)). Captive cotton-top tamarins, a New World nonhuman primate species, develop spontaneous, often chronic, colitis that clinically and histolgocially resembles ulcerative colitis in humans (Madara, J. L. et al., Gastroenterology, 88: 13-19 (1985)). The tamarin model and other animal models of gastrointestinal inflammation using BALB/c mice (a (DSS)-induced inflammation model; DSS, dextran sodium sulfate). IL-10 knockout mice which develop intestinal lesions similar to those of human inflammatory bowel disease have also been described (Strober, W. and Ehrhardt, R. O., Cell, 75: 203-205 (1993)).
According to the method, an inhibitor can be administered to an individual (e.g., a human) alone or in conjunction with another agent, such as an additional pharmacologically active agent (e.g., sulfasalazine, an antiinflammatory compound, or a steroidal or other nonsteroidal antiinflammatory compound). A compound can be administered before, along with or subsequent to administration of the additional agent, in amounts sufficient to reduce or prevent MAdCAM-mediated binding to a ligand for MAdCAM-1, such as human xcex14xcex27.
An effective amount of the active ingredient can be administered by an appropriate route in a single dose or multiple doses. An effective amount is a therapeutically effective amount sufficient to achieve the desired therapeutic and/or prophylactic effect (such as an amount sufficient to reduce or prevent MAdCAM-mediated binding to a MAdCAM ligand, thereby inhibiting leukocyte adhesion and infiltration and associated cellular responses. Suitable dosages of active ingredient of the present invention for use in therapy, diagnosis or prophylaxis, can be determined by methods known in the art and can be dependent, for example, upon the individual""s age, sensitivity, tolerance and overall well-being.
The active ingredient of the present invention or pharmaceutically acceptable salts thereof may be administered either orally or parenterally, and it may be used as a suitable pharmaceutical preparation, for example, a tablet, a granule, a capsule, a powder, an injection, and an inhalation by a conventional process.
The dose of the active ingredient of the present invention or a pharmaceutically acceptable salt thereof varies depending on an administration method, age, body weight, and state of a patient, but, in general, the daily dose is preferably about 0.1 to 100 mg/kg/day, particularly preferably 1 to 100 mg/kg/day.
Pharmaceutical Compositions
As indicated previously, the active ingredient of formula [I] can be formulated into pharmaceutical compositions. In determining when a compound of formula [I] is indicated for the treatment of a given disease, the particular disease in question, its severity, as well as the age, sex, weight, and condition of the subject to be treated, must be taken into consideration and this perusal is to be determined by the skill of the attendant physician.
For medical use, the amount of a compound of Formula [I] required to achieve a therapeutic effect will, of course, vary both with the particular compound, the route of administration, the patient under treatment, and the particular disorder or disease being treated. A suitable daily dose of a compound of Formula [I], or a pharmaceutically acceptable salt thereof, for a mammalian subject suffering from, or likely to suffer from, any condition as described herein before is 0.1 mg to 100 mg of the compound of formula [I], per kilogram body weight of the (systemic) mammalian subject. In the case of systemic administration, the dose may be in the range of 0.5 to 100 mg of the compound per kilogram body weight, the most preferred dosage being 0.5 to 50 mg/kg of mammal body weight administered two to three times daily. In the case of topical administration, e.g., to the skin or eye, a suitable dose may be in the range of 0.1 xcexcg to 100 xcexcg of the compound per kilogram, typically about 0.1 xcexcg/kg.
In the case of oral dosing, a suitable dose of a compound of Formula [I], or a physiologically acceptable salt thereof, may be as specified in the preceding paragraph, but preferably is from 1 mg to 50 mg of the compound per kilogram, the most preferred dosage being from 5 mg to 25 mg/kg of mammal body weight, for example, from 1 to 10 mg/kg. Most preferably, a unit dosage of an orally administrable composition encompassed by the present invention contains less than about 1.0 g of a formula [I] compound.
It is understood that the ordinarily skilled physician or veterinarian will readily determine and prescribe the effective amount of a compound of Formula [I] to prevent or arrest the progress of the condition for which treatment is administered. In so proceeding, the physician or veterinarian could employ relatively low doses at first, subsequently increasing the dose until a maximum response is obtained.
The compounds and compositions of the present invention can be administered to patients suffering from a condition listed herein in an amount which is effective to fully or partially alleviate undesired symptoms of the condition. The symptoms may be caused by inappropriate cell adhesion or cell activation to release proinflammatory mediators mediated by xcex14xcex27 integrins. Such inappropriate cell adhesion or signal transduction would typically be expected to occur as a result of increased VCAM-1 and/or MAdCAM expression on the surface of endothelial cells. Increased VCAM-1, MAdCAM and/or CS-1 expression can be due to a normal inflammation response or due to abnormal inflammatory states. In either case, an effective dose of a compound of the invention may reduce the increased cell adhesion due to increased VCAM-1 and/or MAdCAM expression by endothelial cells. Reducing the adhesion observed in the disease state by 50% can be considered an effective reduction in adhesion. More preferably, a reduction in ex vivo adhesion by 90%, is achieved. Most preferably, adhesion mediated by VCAM-1, MAdCAM and/or CS-1 interaction is abolished by an effective dose. Clinically, in some instances, effect of the compound can be observed as a decrease in white cell infiltration into tissues or a site of injury. To achieve a therapeutic effectiveness, then, the compounds or compositions of the present invention are administered to provide a dose effective to reduce or eliminate inappropriate cell adhesion or inappropriate cell activation to alleviate undesired symptoms.
While it is possible for an active ingredient to be administered alone, it is preferable to present it as a pharmaceutical formulation comprising a compound of Formula [I] and a pharmaceutically acceptable carrier thereof. Such formulations constitute a further feature of the present invention.
The formulations, both for human and veterinary medical use, of the present invention comprise an active ingredient of Formula [I], in association with a pharmaceutically acceptable carrier thereof and optionally other therapeutic ingredients), which are generally known to be effective in treating the disease or condition encountered. The carrier(s) must be xe2x80x9cacceptablexe2x80x9d in the sense of being compatible with the other ingredients of the formulations and not deleterious to the recipient thereof.
The formulations include those in a form suitable for oral, pulmonary, ophthalmic, rectal, parenteral (including subcutaneous, intramuscular, and intravenous), intraarticular, topical, nasal inhalation (e.g., with an aerosol) or buccal administration. Such formulation are understood to include long-acting formulations known in the art. Oral and parenteral administration are preferred modes of administration.
The formulations may conveniently be presented in unit dosage form and may be prepared by any of the methods well known in the art of pharmacy. All methods may include the step of bringing the active ingredient into association with the carrier which constitutes one or more accessory ingredients. In general, the formulations are prepared by uniformly and intimately bringing the active ingredient into association with a liquid carrier or a finely divided solid carrier or both, and then, if necessary, shaping the product into the desired form.
Formulations of the present invention suitable for oral administration may be in the form of discrete units such as capsules, cachets, tablets, or lozenges, each containing a predetermined amount of the active ingredient in the form of a powder or granules; in the form of a solution or suspension in an aqueous liquid. Formulations for other uses could involve a nonaqueous liquid; in the form of an oil-in-water emulsion or a water-in-oil emulsion; in the form of an aerosol; or in the form of a cream or ointment or impregnated into a transdermal patch for use in administering the active ingredient transdermally, to a patient in need thereof. The active ingredient of the present inventive compositions may also be administered to a patient in need thereof in the form of a bolus, electuary, or paste.
The practitioner is referred to xe2x80x9cRemington: The Science and Practice of Pharmacy,xe2x80x9d 19th Edition, c. 1995 by the Philadelphia College of Pharmacy and Science, as a comprehensive tome on pharmaceutical preparations.
According to the present invention, the novel compound [I] can be prepared by the following methods.
Method A 
(wherein R4a is an ester group, and other symbols are the same as defined above)
The compound of the formula [I] or a pharmaceutically acceptable salt thereof may be prepared by:
(1) condensing a compound of the formula [II] a salt thereof or a reactive derivative thereof with a compound of the formula [III] or a salt thereof,
(2) converting the ester group of the compound of the formula [Ia] into a carboxyl group, if desired, and
(3) converting the carboxyl group of the resulting compound into an ester group, an amide group, a tetrazolyl group or a pharmaceutically acceptable salt thereof, if further desired.
A salt of the compound [II] and/or [III] includes, for example, a salt with an inorganic acid (e.g., trifluoroacetate, hydrochloride, sulfate), a salt with an inorganic base (e.g., an alkali metal salt such as a sodium salt or a potassium salt, an alkaline earth metal salt such has a barium salt or calcium salt).
(1) The condensation reaction can be carried out by a conventional method for a usual amide bond synthesis.
The condensation reaction of the compound [II] or a salt thereof with the compound [III] or a salt thereof is carried out in the presence of a condensing reagent with or without a base in a suitable solvent or without a solvent. The condensing reagent can be selected from any one which can be used for a conventional amide bond synthesis, for example, BOP-Cl, BOP reagent, DCC, EDC or CDI.
The base can be selected from an organic base (e.g., DIEA, DMAP, DBU, Et3N), an alkali metal hydride (e.g., NaH, LiH), an alkali metal carbonate (e.g., Na2CO3, K2CO3), an alkali metal hydrogen carbonate (e.g., NaHCO3, KHCO3), an alkali metal amide (e.g., NaNH2), an alkali metal alkoxide (e.g., NaOMe, KOMe), a lower alkyl alkali metal salt (e.g., n-BuLi, t-BuLi), an alkali metal hydroxide (e.g., NaOH, KOH), an alkaline earth metal hydroxide (e.g., Ba(OH)2), and the like.
The solvent can be selected from any one which does not disturb the condensation reaction, for example, CH2Cl2, THF, DMF or a mixture thereof. The reaction is carried out at a temperature of 0xc2x0 C. to room temperature, preferably at room temperature.
The condensation reaction of the compound [III] or a salt thereof with the reactive derivative of the compound [II], for example, with an acid halide (e.g., an acid chloride), a reactive ester (e.g., an ester with p-nitrophenol), an anhydride thereof, a mixed anhydride with other carboxylic acid (e.g., a mixed anhydride with acetic acid), and the like, is carried out in the presence of a base or without a base in a solvent or without a solvent.
The base can be selected from an organic base (e.g., DIEA, DMAP, DBU, Et3N), an alkali metal hydride (e.g., NaH, LiH), an alkali metal carbonate (e.g., Na2CO3, K2CO3), an alkali metal hydrogen carbonate (e.g., NaHCO3, KHCO3), an alkali metal amide (e.g., NaNH2), an alkali metal alkoxide (e.g., NaOMe, KOMe), a lower alkylalkali metal salt (e.g., n-BuLi, t-BuLi), an alkali metal hydroxide (e.g., NaOH, KOH), an alkaline earth metal hydroxide (e.g., Ba(OH)2), and the like.
The solvent can be selected from any one which does not disturb the condensation reaction, for example, CH2Cl2, C2H4Cl2, Et2O, THF, DMF, CH3CN, DMSO, benzene, toluene or a mixture thereof. The reaction is carried out at a temperature of xe2x88x9230xc2x0 C. to 100xc2x0 C.
(2) The conversion of the ester group into a carboxyl group can be carried out by a conventional method, which is selected according to the type of the ester group to be removed, for example, hydrolysis using a base (e.g., LiOH, NaOH) or an acid (e.g., HCl), treatment with an acid (e.g., TFA), catalytic reduction using a catalyst (e.g., palladium on activated carbon) and the like. The ester group can be selected from a conventional ester, for example, a lower alkyl ester, a lower alkenyl ester, a lower alkynyl ester, an aryl-lower alkyl ester (e.g., benzyl ester), an aryl ester (e.g., phenyl ester) and the like.
(3) The conversion of the carboxyl group into an ester group, an amide group or tetrazolyl group or conversion of the compound into a pharmaceutically acceptable salt thereof can be carried out by a conventional method. Particularly, the conversion of the carboxyl group into an ester group or an amide group can be carried out in a similar manner as described in Method A-(1). The conversion of the carboxyl group into tetrazolyl group is detailed in Procedure N below.
Method B 
(wherein X1 is a leaving group and other symbols are the same as defined above.)
The compound of the formula [I] can be prepared by:
(1) reacting a compound of the formula [IV] with a compound of the formula [V],
(2) converting the ester group of the compound of the formula [Ia] into a carboxyl group, if desired, and
(3) converting the carboxyl group of the resulting compound into an ester group, an amide group, a tetrazolyl group or a pharmaceutically acceptable salt thereof, if further desired.
Examples of the leaving group X1 may be a halogen atom and a trifluoromethanesulfonyloxy group.
(1) The coupling reaction can be carried out by a conventional aryl coupling method, e.g., Suzuki coupling method (for reference of Suzuki coupling method: (a) Suzuki et al., Synth. Commun. 1981, 11, 513, (b) Suzuki, Pure and Appl. Chem. 1985, 57, 1749-1758, (c) Suzuki et al., Chem. Rev. 1995, 95, 2457-2483, (d) Shieh et al., J. Org. Chem. 1992, 57, 379-381), (e) Martin et al., Acta Chemica Scandinavica, 1993, 47, 221-230.)
The coupling reaction can be carried out, for example, at a temperature of room temperature to 100xc2x0 C., preferably at a temperature of 80xc2x0 C. to 100xc2x0 C., in the presence of tetrakis(triphenylphosphine)palladium and a base (e.g., an inorganic base such as K2CO3) in an organic solvent. The organic solvent can be selected from any one which does not disturb the coupling reaction, for example, toluene, DME, DMF, H2O or a mixture thereof.
(2) The conversion of ester group into carboxyl group can be carried out according to Method A-(2).
(3) The conversion of carboxyl group into ester group or amide group, a tetrazolyl group or pharmaceutically acceptable salt can be carried out according to Method A-(3).
Method C 
(wherein symbols are the same as defined above.)
The compound of the formula [I] can be also prepared by:
(1) converting the compound [IV] to the corresponding organotin compound (e.g., the compound of the formula [VII]),
(2) reacting the compound [VII] with a compound of the formula [VIII]:
R6xe2x80x94Xxe2x80x83xe2x80x83[VIII]
xe2x80x83wherein X is a leaving group and R6 is the same as defined above,
(3) converting the ester group of the compound of the formula [Ia] into a carboxyl group, if desired, and
(4) converting the carboxyl group of the resulting compound into an ester group, an amide group, a tetrazolyl group or a pharmaceutically acceptable salt thereof, if further desired.
Examples of the leaving group X is a halogen atom and a trifluoromethanesulfonyloxy group.
(1) The conversion of the compound [IV] to the organotin compound [VII] can be carried out, for example, by reacting the compound [IV] with hexaalkylditin (e.g., hexamethylditin) at a temperature of room temperature to 150xc2x0 C., preferably at a temperature of 80xc2x0 C. to 110xc2x0 C., in the presence of tetrakis(triphenylphosphine)palladium and an additive (e.g., LiCl) in an organic solvent. The organic solvent can be selected from any one which does not disturb the coupling reaction, for example, dioxane, toluene, DME, DMF, H2O or a mixture thereof.
(2) The coupling reaction can be carried out by a conventional aryl coupling method, e.g., Stille coupling method (for reference of Stille coupling method: Stille et al., Angew. Chem. Int. Ed. Engl., 25, 508 (1986))
The coupling reaction can be carried out, for example, at a temperature of room temperature to 150xc2x0 C., preferably at a temperature of 80xc2x0 C. to 120xc2x0 C., in the presence of tetrakis(triphenylphosphine)palladium in an organic solvent. The organic solvent can be selected from any one which does not disturb the coupling reaction, for example, toluene, DME, DMF, H2O or a mixture thereof.
(3) The conversion of ester group into carboxyl group can be carried out according to Method A-(2).
(4) The conversion of carboxyl group into ester group or amide group, a tetrazolyl group or pharmaceutically acceptable salt can be carried out according to Method A-(3).
The compound [IV] may be prepared by condensing the compound of the formula [IIa]: 
wherein Y is a halogen atom and the other symbols are the same as defined above, with the compound of the formula [IIIa]: 
wherein the symbols are the same as defined above or a salt thereof by the conventional method for the usual peptide synthesis as described above for the condensation reaction of the compound [III] or a salt thereof with the reactive derivative of the compound [II] (e.g., an acid halide).
The compound [IV] may be also prepared by:
(1) condensing the compound [IIa] with the compound of the formula [IIIb]: 
wherein the symbols are the same as defined above or a salt thereof by the similar manner as described above,
(2) converting the hydroxyl group of the resulting compound into a leaving group by the conventional method. For example, the conversion of the hydroxy group into trifluoromethanesulfonyloxy group can be carried out by using triflic anhydride at 0xc2x0 C. in the presence of a base(e.g., pyridine, NEt3, DIEA) in an organic solvent (e.g., CH2Cl2, THF or a mixture thereof).
The compound [III] may be prepared by:
(1) condensing the compound of the formula [VIa]: 
xe2x80x83wherein P is a protecting group for an amino group and other symbols are the same as defined above with the compound [V] by a conventional aryl coupling method which is well known as Suzuki coupling method,
(2) removing the protecting group for the amino group of the resulting compound.
The protecting group for the amino group can be selected from a conventional protecting group for an amino group, for example, a substituted or unsubstituted aryl-lower alkoxycarbonyl group (e.g., benzyloxycarbonyl group, p-nitrobenzyloxycarbonyl group), a lower alkoxycarbonyl group (e.g., tert-butoxycarbonyl group) and the like.
The removal of the protecting group for the amino group can be carried out by a conventional method, which is selected according to the type of the protecting group to be removed, for example, catalytic reduction using a catalyst (e.g., palladium on activated carbon), treatment with an acid (e.g., TFA) and the like.
The condensation reaction can be carried out in a similar manner as described for the coupling reaction of the compound [IV] and [V].
The compound [VIa] wherein X1 is trifluoromethanesulfonyloxy group may be prepared by reacting the compound of the formula [VIb]: 
wherein the symbols are the same as defined above with triflic anhydride in a similar manner as described for the preparation of the compound [IV].
The compound [V] may be prepared by a conventional method (e.g., reference (a) Kuivila et al., J. Am. Chem. Soc., 1961, 83, 2159; (b) Gerrard, The Chemistry of Boron; Academic Press: New York, 1961; (c) Muetterties, The Chemistry of Boron and its Compounds: Wiley: New York, 1967; (d) Alamansa et al., J. Am. Chem. Soc., 1994, 116, 11723-11736):
(1) reacting a substituted or unsubstituted aryl lithium or a substituted or unsubstituted heteroaryl lithium with trimethyl borate at a temperature of xe2x88x92100xc2x0 C. to room temperature in an organic solvent (e.g., diethyl ether, THF or the mixture thereof), and
(2) hydrolyzing the resulting compound by a conventional method.
The hydrolysis can be carried out at room temperature in an organic solvent (e.g., diethyl ether, THF or the mixture thereof) in the presence of mild acid (e.g., AcOH or citric acid) and water.
The desired compound [I] of the present invention can be converted to each other. Such conversion of the present compound [I] into the other compound [I] may be carried out in an organic solvent by selecting one of the following procedures (Procedure A to K) according to the type of the substituent thereof. The organic solvent can be selected from any one which does not disturb the said procedure.
Procedure A: Reduction of Carbonyl Group
The compound [I] wherein R1, R2, R3, R5 or the substituent of the R6 group is a hydroxy-lower alkyl group such as a hydroxymethyl group or a group of the formula: lower alkyl-CH(OH)xe2x80x94 can be prepared by the reduction of the compound [I] wherein the corresponding R1, R2, R3, R5 or the substituent of the R6 group is a carboxyl group, a formyl group or a group of the formula: lower alkyl-COxe2x80x94. The reduction reaction can be carried out by a conventional method using a reducing agent such as borane, alkali metal borohydride (e.g., sodium borohydride) and the like at a temperature of 0xc2x0 C. to room temperature in an organic solvent, e.g., methanol, ethanol, THF or the mixture thereof.
Procedure B: Oxidation of Formyl Group
The compound [I] wherein R1, R2, R3, R5 or the substituent of the R6 group is a carboxyl group can be prepared by the oxidation of the compound [I] wherein the corresponding R1, R2, R3, R5 or the substituent of the R6 group is a formyl group. The oxidation reaction can be carried out by a conventional method using an oxidizing agent, e.g., KMnO4 and the like at a temperature of 0xc2x0 C. to 50xc2x0 C., preferably at a temperature of 30xc2x0 C. to 50xc2x0 C., in an organic solvent, e.g., acetone, H2O or the mixture thereof.
Procedure C: Reduction of Nitro Group
The compound [I] wherein R1, R2, R3, R5 or the substituent of the R6 group is an amino group or has an amino group can be prepared by the reduction of the compound [I] wherein the corresponding R1, R2, R3, R5 or the substituent of the R6 group is a nitro group or has a nitro group. The reduction reaction can be carried out by a conventional method, e.g., 1) a catalytic reduction using a catalyst such as Raney-nickel or a palladium on activated carbon and the like under a hydrogen atmosphere at room temperature in an organic solvent, e.g., methanol, H2O or the mixture thereof, 2) chemical reduction using metal and inorganic acid, such as Fe/HCl, Sn/HCl, SnCl2/HCl and the like, or 3) reduction with a reducing agent, such as Na2S2O4, in a suitable solvent, e.g., methanol, ethanol, H2O or the mixture thereof or without a solvent at a temperature of 0xc2x0 C. to 80xc2x0 C.
Procedure D: Removal of Protecting Group
(D-1) The compound [I] wherein R1, R2, R3, R5 or the substituent of the R6 group is an amino group or has an amino group can be prepared by the deprotection of the amino group of the compound [I] wherein the corresponding R1, R2, R3, R5 or the substituent of the R6 groups is an N-protected amino group or has an N-protected amino group and the protecting group is a conventional protecting group for an amino group, e.g., benzyloxycarbonyl group, tert-butoxycarbonyl group, 9-fluorenylmethoxycarbonyl group, allyl group and the like. The deprotection reaction can be carried out by a conventional method, which is selected according to the type of the protecting group to be removed, e.g., 1) catalytic reduction using a catalyst such as palladium on activated carbon under a hydrogen atmosphere, 2) a treatment with an acid such as hydrogen chloride or TFA, 3) a treatment with an amine such as piperidine, 4) a treatment with a catalyst such as Wilkinson""s catalyst, at room temperature or with heating in an organic solvent, e.g., CH2Cl2, THF, MeOH, EtOH and MeCN, or without an organic solvent.
(D-2) The compound [I] wherein R1, R2, R3, R5 or the substituent of the R6 group is a sulfamoyl group can be prepared by the deprotection of the compound [I] wherein the corresponding R1, R2, R3, R5 or the substituent of the R6 group is an N-protected sulfamoyl group and the protecting group is a conventional protecting group for a sulfamoyl group, e.g., tert-butyl group and the like. The deprotection reaction can be carried out by a conventional method, which is selected according to the type of the protecting group to be removed, e.g., a treatment with an acid such as TFA at a room temperature in an organic solvent, e.g., CH2Cl2, or without an organic solvent.
(D-3) The compound [I] wherein R1, R2, R3, R4, R5 or the substituent of the R6 group is a carboxyl group or has a carboxyl group can be prepared by the deprotection of the compound [I] wherein the corresponding R1, R2, R3, R4, R5 or the substituent of the R6 group is a protected carboxyl group or has a protected carboxyl group and the protecting group is a conventional protecting group for a carboxyl group, e.g., a lower alkyl group, a lower alkenyl group, a lower alkynyl group, an aryl-lower alkyl group, an aryl group and the like. The deprotection reaction can be carried out by a conventional method, which is selected according to the type of the protecting group to be removed, for example, hydrolysis using a base (e.g., NaOH, LiOH, KOH) or an acid (e.g., hydrochloric acid) treatment with an acid (e.g., TFA), catalytic reduction using a catalyst (e.g., palladium on activated carbon) and the like, at room temperature in an organic solvent (e.g., MeOH, EtOH or THF) or without an organic solvent.
(D-4) The compound [I] wherein R1, R2, R3, R5 or the substituent of the R6 group is a hydroxyl group or has a hydroxyl group can be prepared by the deprotection of the compound [I] wherein the corresponding R1, R2, R3, R5 or the substituent of the R6 group is a protected hydroxyl group or has a protected hydroxyl group and the protecting group is a conventional protecting group for a hydroxyl group, e.g., a methyl group, methoxymethyl group, tetrahydropyranyl group and the like. The deprotection reaction can be carried out by a conventional method, which is selected according to the type of the protecting group to be removed, for example, a treatment with BBr3 for the demethylation of a methoxy group, and a treatment with HCl at a temperature of xe2x88x9278xc2x0 C. to room temperature in an organic solvent, e.g., CH2Cl2 and MeOH for removal of methoxymethyl group.
Procedure E: Acylation of Amino Group
(E-1) The compound [I] wherein R1, R2, R3, R5 or the substituent of the R6 group is an N-acylamino group, e.g., a lower alkanoylamino group, a lower alkoxycarbonylamino group, an arylcarbonylamino group, a chlorosulfonylcarbamoylamino group (such as 3-chlorosulfonylureido group), a lower alkyl carbamoylamino group (such as 3-(lower alkyl)ureido group), a substituted or unsubstituted arylcarbamoyl amino group (such as 3-(substituted or unsubstituted aryl)ureido group), a (substituted or unsubstituted lower alkyl)thiocarbamoylamino group (such as 3-(lower alkyl)thioureido group, 3-(phenyl-lower alkyl)thioureido group) can be prepared by the N-acylation of the compound [I] wherein the corresponding R1, R2, R3, R5 or the substituent of the R6 group is an amino group. The N-acylation reaction can be carried out by a conventional method using 1) an acylating reagent, e.g., a lower alkanoyl halide, a lower alkanoic acid anhydride, a lower alkyl halogenoformate such as a lower alkyl chloroformate, an aryl carbonyl halide, a chlorosulfonyl isocyanate, a lower alkyl isocyanate, a substituted or unsubstituted aryl isocyanate or a lower alkyl isocyanate, or 2) when preparing a lower alkoxycarbonylamino group, a (lower alkyl)carbamoylamino group, a substituted or unsubstituted arylcarbamoyl amino group, a (substituted or unsubstituted lower alkyl)thiocarbamoylamino group, a condensing reagent, e.g., CDI, thioCDI, and a requisite amine or alcohol, at a temperature of 0xc2x0 C. to 100xc2x0 C., preferably at a temperature of room temperature to 90xc2x0 C., with a base (e.g., DIEA, DMAP, pyridine, NaHCO3, Na2CO3, KHCO3, K2CO3) or without a base in an organic solvent (e.g., THF, CH3CN, CH2Cl2, DMF, toluene, acetone and the mixture thereof).
(E-2) The compound [I] wherein R1, R2, R3, R5 or the substituent of the R6 group is an N-(lower alkylsulfonyl)amino group (e.g., methanesulfonylamino group), an N-(substituted or unsubstituted arylsulfonyl)amino group (e.g., p-toluenesulfonylamino group, benzenesulfonylamino group) or an N-(substituted or unsubstituted heteroarylsulfonyl)amino group (e.g., quinolinosulfonylamino group) can be prepared by the N-sulfonylation of the compound [I] wherein the corresponding R1, R2, R3, R5 or the substituent of the R6 group is an amino group. The N-sulfonylation reaction can be carried out by a conventional method using a lower alkylsulfonyl halide or a substituted or unsubstituted arylsulfonyl halide or a substituted or unsubstituted heteroarylsulfonyl halide in the presence of a base (e.g., pyridine, DMAP, Et3N, DIEA, NaHCO3, KHCO3, Na2OC3, K2CO3) at a temperature of 0xc2x0 C. to room temperature, preferably at room temperature, in an organic solvent (e.g., CH2Cl2, THF, DMF, CH3CN, toluene, acetone and the mixture thereof).
(E-3) The compound [I] wherein R1, R2, R3, R5 or the substituent of the R6 group is a ureido group can be prepared by the hydrolysis of the compound [I] wherein the corresponding R1, R2, R3, R5 or the substituent of the R6 group is a 3-chlorosulfonylureido group. The hydrolysis can be carried out using a base (e.g., LiOH, NaOH and the like) or an acid (e.g., HCl) at room temperature in a suitable solvent (e.g., THF, CH3CN, H2O) or a mixture thereof.
Procedure F: Alkylation of Hydroxyl Group
The compound [I] wherein R1, R2, R3, R5 or the substituent of the R6 group is a substituted or unsubstituted lower alkoxy group, e.g., a substituted or unsubstituted hetero-cycloalkyl-lower alkoxy group (such as a substituted or unsubstituted piperidyl-lower alkoxy group, or a substituted or unsubstituted pyrrolidinyl-lower alkoxy group), an aryl-lower alkoxy group, a heteroaryl-lower alkoxy group (such as a pyridyl-lower alkoxy group, a substituted or unsubstituted thiazolyl-lower alkoxy group, a substituted or unsubstituted isoxazolyl-lower alkoxy group, a substituted or unsubstituted thienyl-lower alkoxy group), a lower alkoxycarbonyl-lower alkoxy group, a carboxy-lower alkoxy group, a hydroxy-lower alkoxy group, a cyano-lower alkoxy group or a lower alkoxy group, can be prepared by the alkylation of the compound [I] wherein the corresponding R1, R2, R3, R5 or the substituent of the R6 group is a hydroxy group, followed by the deprotection of the protecting group for carboxyl group or hydroxyl group by a conventional method, if desired. The alkylation reaction can be carried out using a halogenated lower alkane not having a substituent (e.g., methyl iodide) or that having a substituent such as a substituted or unsubstituted aryl group (e.g., unsubstituted aryl-lower alkyl halide such as benzyl bromide), a substituted or unsubstituted heteroaryl group (e.g., substituted or unsubstituted heteroaryl-lower alkyl halide such as pyridylmethyl bromide, isoxazolylmethyl bromide, thiazolylmethyl bromide), a heterocycloalkyl group (e.g., substituted heterocycloalkyl-lower alkyl halide such as N-lower alkylpyrrolidinyl-lower alkyl bromide, N-lower alkylpiperidyl-lower alkyl bromide), a lower alkoxycarbonyl group (e.g., halogenoalkanoic acid lower alkyl ester such as methyl bromoacetate) or a cyano group (e.g., bromoacetonitrile) in the presence of a base (e.g., Et3N, DIEA, NaHCO3, KHCO3, Na2CO3, K2CO3, KHCO3, CsCO3) at a temperature of room temperature to 50xc2x0 C. in an organic solvent (e.g., CH2Cl2, THF, DMF, CH3CN, toluene).
The alkylation reaction can be also carried out by using a conventional alkylation method such as Mitsunobu Reaction (for reference of Mitsunobu reaction: (a) Mitsunobu, Synthesis, 1-28, (1981), (b) Hughes, Organic Reactions, 42, 335 (1992), (c) Mitsuhashi et al., J. Am. Chem. Soc., 94, 26 (1972)).
Procedure G: Halogenation of Hydroxyl Group
The compound [I] wherein R1, R2, R3, R5 or the substituent of the R6 group is a halogenated lower alkyl group can be prepared by the halogenation of the compound [I] wherein the corresponding R1, R2, R3, R5 or the substituent of the R6 group is a hydroxyl lower alkyl group. The halogenation reaction can be carried out by the conventional method using, for example, a combination of tetrahalomethane (e.g., CBr4) and triphenylphosphine at a room temperature in an organic solvent (e.g., CH2Cl2).
Procedure H: Conversion of Halogenated Alkyl Group to Alkoxy Alkyl Group
The compound [I] wherein R1, R2, R3, R5 or the substituent of the R6 group is a lower alkoxy-lower alkyl group can be prepared by reacting the compound [I] wherein the corresponding R1, R2, R3, R5 or the substituent of the R6 group is a halogenated lower alkyl group with an alkali metal lower alkoxide (e.g., sodium methoxide) at room temperature in an organic solvent (e.g., DMF, THF, CH3CN).
Procedure I: Conversion of Carboxyl Group into Carbamoyl Group
The compound [I] wherein R1, R2, R3, R4, R5 or the substituent of the R6 group is a substituted or unsubstituted carbamoyl group (e.g., an N-lower alkylcarbamoyl group, an N,N-(lower alkyl)(lower alkyl)carbamoyl group, an N-(hydroxy-lower alkyl)carbamoyl group, an N-(morpholino-lower alkyl)carbamoyl group, an N-(aryl-lower alkyl)carbamoyl group, an N-(lower alkanesulfonyl)carbamoyl group, a hydroxycarbamoyl group, a carbamoyl group) can be prepared by condensing the compound [I] wherein the corresponding R1, R2, R3, R4, R5 or the substituent of the R6 group is a carboxyl group with a substituted or unsubstituted amine (e.g., a lower alkylamine, an N,N-(lower alkyl)(lower alkyl)amine, a (hydroxy-lower alkyl)amine, a (morpholino-lower alkyl)amine, an (aryl-lower alkyl)amine, hydroxyamine, ammonia) or a lower alkanesulfonamide.
The condensation reaction can be carried out by the conventional method for a usual peptide synthesis as described for the condensing reaction of the compound [II] and [III].
Procedure J: Reductive Alkylation
(J-1) The compound [I] wherein R1, R2, R3, R5 or the substituent of the R6 group is an amino-lower alkyl group, a lower alkyl amino-lower alkyl group or an arylamino-lower alkyl group can be prepared by the reductive alkylation of the corresponding ammonia, lower alkyl amine or aryl amine with the compound [I] wherein the corresponding R1, R2, R3, R5 or the substituent of the R6 group is a formyl group. The reductive alkylation reaction can be carried out by the conventional method using a reductive agent (e.g., sodium cyanoborohydride) and an acid (e.g., HCl) at room temperature in an organic solvent (e.g., MeOH, THF, dioxane, or the mixture thereof).
(J-2) The compound [I] wherein R1, R2, R3, R5 or the substituent of the R6 group is an N,N-dimethylamino group can be prepared by the reductive alkylation of the compound [I] wherein the corresponding R1, R2, R3, R5 or the substituent of the R6 group is an amino group. The reductive alkylation can be carried out by the conventional method using formaldehyde, a reducing agent (e.g., sodium cyanoborohydride) and an acid (e.g., HCl) at room temperature in an organic solvent (e.g., MeOH, EtOH, THE, dioxane) or H2O or the mixture thereof.
Procedure K: Wittig Reaction
The compound [I] wherein R1, R2, R3, R5 or the substituent of the R6 group is a lower alkokycarbonyl-ethenyl group can be prepared by the Wittig reaction of the compound [I] wherein the corresponding R1, R2, R3, R5 or the substituent of the R6 group is a formyl group. The Wittig reaction can be carried out by the conventional method using, for example, (triphenylphosphoranylidene)acetic acid lower alkyl ester at a temperature of 50xc2x0 C. to 100xc2x0 C. in an organic solvent (e.g., toluene, THF).
Procedure L: Conversion of Halogenated Alkyl Group to Amino Alkyl Group
The compound [I] wherein R1, R2, R3, R5 or the substituent of the R6 group is a lower alkyl group which is substituted by a substituted or unsubstituted amino group, a substituted or unsubstituted piperidinyl group, a substituted or unsubstituted morpholino group, a thiomorpholino group which may be oxidized, a substituted or unsubstituted piperazinyl group, or a substituted or unsubstituted pyrrolidinyl group can be prepared by reacting the compound [I] wherein the corresponding R1, R2, R3, R5 or the substituent of the R6 group is a halogenated lower alkyl group with a requisite amine at room temperature or under cooling in an organic solvent (e.g., DMF, THF, CH2Cl2) or without a solvent, with or without a base such as Et3N, DIEA.
In particular, the compound [I] wherein R1 and R5 are hydrogen atoms, R2 and R3 are halogen atoms, and R6 is a phenyl group substituted by a lower alkoxy group and a lower alkyl group which is substituted by a group selected from the group consisting of a substituted or unsubstituted amino group, a substituted or unsubstituted piperidinyl group, a substituted or unsubstituted morpholino group, a substituted or unsubstituted piperazinyl group and a substituted or unsubstituted pyrrolidinyl group can be prepared by reacting the compound [I] wherein R1 and R5 are hydrogen atoms, R2 and R3 are halogen atoms, and R6 is a phenyl group substituted by a lower alkoxy group and a halogeno-lower alkyl group with a requisite amine such as a substituted or unsubstituted ammonia, a substituted or unsubstituted piperidine, a substituted or unsubstituted morpholine, a substituted or unsubstituted piperazine and a substituted or unsubstituted pyrrolidine. The reaction can be carried out as described above.
Procedure M: Conversion of Carbonyl Group to Thiocarbonyl Group
The compound wherein Z is sulfur atom can be prepared by reacting the compound [I] wherein Z is oxygen atom with Lawesson""s reagent in a suitable organic solvent (e.g., toluene, xylene) at a temperature of 50xc2x0 C. to 150xc2x0 C.
Procedure N: Conversion of Carboxyl Group to Tetrazoly Group
The compound [I] wherein R4 is tetrazolyl group can be prepared from the compound [I] wherein R4 is carboxyl group following the procedure described in the J. Med. Chem., 41, 1513-1518, 1998. The procedure can be summarized in the following scheme: 
Procedure O: Conversion of Carboxyl Group to Alkoxycarbonyl Group
The compound [I] wherein R1, R2, R3, R4, R5 or the substituent of the R6 group is a substituted or unsubstituted lower alkoxycarbonyl group can be prepared by condensing the compound [I] wherein the corresponding R1, R2, R3, R4, R5 or the substituent of the R6 group is a carboxyl group with a substituted or unsubstituted alcohol (e.g., a halogeno-lower alcohol, pyridyl-lower alcohol, a (lower alkylamino)-lower alcohol, a lower alkoxy-lower alcohol).
The condensation reaction can be carried out by the conventional method for a usual ester synthesis as described for Method A-(3).
Procedure P: Reduction of Hydroxyl Group
The compound [I] wherein R1, R2, R3, R5 or the substituent of the R6 group is a lower alkyl group can be prepared by reducing the compound [I] wherein the corresponding R1, R2, R3, R5 or the substituent of the R6 group is a hydroxy-lower alkyl group. The reduction can be carried out using a reducing reagent such as a silane compound (e.g., Et3SiH) in the presence of Lewis acid (e.g., BF3, TiCl4) in a suitable organic solvent (e.g., MeCN, CH2Cl2, THF) at a temperature of 0xc2x0 C. to xe2x88x9278xc2x0 C.
Procedure Q: Halogenation of Phenyl Group
The compound [I] wherein R6 is a substituted or unsubstituted halogeno-phenyl group can be prepared by reacting the compound [I] wherein R6 is a substituted or unsubstituted phenyl group with halogenating reagent (e.g., Bu4NBr3, 3,5-dichloro-1-fluoropyridinium triflate) in a suitable solvent (e.g., MeCN, CH2Cl2, THF) at room temperature or with heating.
Procedure R: Nitration of Phenyl Group
The compound [I] wherein R6 is a substituted or unsubstituted nitro-phenyl group can be prepared by reacting the compound [I] wherein R6 is a substituted or unsubstituted phenyl group with HNO3 in a suitable solvent (e.g., THF, NeCN, MeOH, EtOH) at a temperature of room temperature to 100xc2x0 C.
Procedure S: Converting Phenyl Group to Carbamoyl-phenyl Group
The compound [I] wherein R6 is a substituted or unsubstituted carbamoyl-phenyl group can be prepared by 1) reacting the compound [I] wherein R6 is a substituted or unsubstituted phenyl group with chlorosulfonyl isocyanate and 2) hydrolyzing the obtained compound. The reaction of the compound [I] and the isocyanate can be carried out in a suitable solvent (e.g., MeCN, CH2Cl2, THF) at a temperature of 0xc2x0 C. to room temperature. The hydrolysis can be carried out with an acid (e.g., HCl, HNO3, H2SO4) in a suitable solvent (e.g., MeCN, H2O) at a temperature of room temperature to 100xc2x0 C.
Procedure T: Conversion of Alkanoyl Group to Imino-alkyl Group
The compound [I] wherein R1, R2, R3, R5 or the substituent of the R6 group is a (hydroxyimino)-lower alkyl or (a lower alkoxyimino)-lower alkyl group can be prepared by reacting the compound [I] wherein the corresponding R1, R2, R3, R5 or the substituent of the R6 group is a lower alkanoyl group with hydroxyamine or a lower alkoxyamine in a suitable solvent such as a lower alcohol (e.g., MeOH, EtOH, PrOH or BuOH) and MeCN, with a base such as alkali metal acetate (e.g., NaOAc) at room temperature or with heating.
Procedure U: Conversion of Halogen Atom to Heterocyclic Group
The compound [I] wherein R1, R2 or R3 is a substituted or unsubstituted heterocyclic group can be prepared by reacting the compound [I] wherein the corresponding R1, R2 or R3 is halogen atom with a (substituted or unsubstituted heterocyclic)boronic acid using a conventional aryl coupling method such as Suzuki Coupling method. The coupling reaction can be carried out following the procedure as describe in Method A.
Procedure V: Oxidation of Sulfur Atom
The compound [I] wherein the substituent of the R6 group is a lower alkylsulfinyl group, a lower alkylsulfonyl group , a thiomorpholino-lower alkyl S-oxide group a thiomorpholino-lower alkyl S,S-dioxide group can be prepared by oxidizing the compound [I] wherein the corresponding substituent of the R6 group is a lower alkylthio group or a thiomorpholino-lower alkyl group with an oxidant such as a peracid (e.g., mCPBA, H2O2, AcOOH, PhCOOOH) in a suitable solvent (e.g., CH2Cl2) at room temperature or under cooling.
Procedure W: Imidation of Hydroxy-lower alkyl
The compound [I] wherein R1, R2, R3 or the substituent of the R6 group is a lower alkyl group which is substituted by succinimido group or 2,5-dioxo-1-imidazolidinyl group optionally substituted by a lower alkyl group can be prepared by the imidation of the compound [I] wherein the corresponding R1, R2, R3 or the substituent of the R6 group is a hydroxy-lower alkyl group. The imidation can be carried out by using a conventional method such as Mitsunobu Reaction (reference of Mitsunobu reaction is made in Procedure F). The reaction can be carried out by reacting the compound [ I] with a di(lower alkyl) azodicarboxylate (e.g., diethyl azodicarboxylate), a tri(lower alkyl)- or triarylphosphine (e.g., triphenylphospphine), and a requisite imide (e.g., succinimide or hydantoin optionally substituted by a lower alkyl group), in a suitable organic solvent (e.g., Et2O and THF) at a temperature of xe2x88x9220xc2x0 C. to 50xc2x0 C.